Process for preparing vaccines based on antigenic ribosomal fractions

Abstract

Vaccines which comprise in association the ribosomal fraction extracted from the bacterias against which a protection is wanted, and the peptidoglycanes extracted from the membranes of at least one of the bacteria previously used.

What is claimed is: 
     
       1. A process for the preparation of an acellular vaccine comprising (1) at least one ribosome extracted from pathogenic bacteria against which protection is desired, said ribosome functioning as an antigenic material and (2) peptidoglycans extracted only from the cell membranes of at least one of the bacteria used in (1), said process comprising (a) extracting ribosomes from said bacteria by (I) cultivating the strain of bacteria corresponding to the desired ribosomal fraction on a growth medium;   (II) decanting the bacterial cells of (I);   (III) grinding said bacterial cells in a buffer solution and eliminating the undestroyed bacteria to form an homogeneous cell macerate;   (IV) ultracentrifuging the homogeneous macerate of said bacteria under an acceleration from about 2×10 4  to 6×10 4  g to form (i) a supernatant phase containing the ribosomes on which impurities of the protein-type are absorbed and (ii) a residue containing all other components;   (V) separating and supernatant phase from said residue;   (VI) ultracentrifuging the separated supernatant phase under an acceleration from about 10 5  to about 2×10 5  g to form a centrifuged residue;   (VII) treating said centrifuged residue with a solution of sodium dodecylsulfate;   (VIII) precipitating sodium dodecylsulfate at low temperature; and   (IX) ultracentrifuging the resulting supernatant solution under an acceleration from about 10 5  to about 2×10 5  g, thus forming a residue containing purified ribosomes;     (b) forming peptidoglycans from the cell membranes of said bacteria by (I) treating the residue of step (a) (IV) (ii) with a saline solution   (II) centrifuging the resulting saline solution-residue mixture to extract the bacterial cell membranes;   (III) digesting the extracted membranes with a proteolytic enzyme to separate membranal glycopeptides, and (IV) centrifuging the membranal glycopeptides, and     (c) mixing the purified ribosomes of (a) (IX) and the membranal glycopeptides of (b) (IV).   
     
     
       2. The process of claim 1, further comprising heating the centrifuged residue of (a) (IV) (I) to 100° C. 
     
     
       3. The process of claim 2 wherein said saline solution of (b) (I) is a 1 M NaCl solution. 
     
     
       4. The process of claim 3 wherein said proteolytic enzyme of (b) (III) is trypsin or chymotrypsin. 
     
     
       5. The process of claim 1 wherein the bacteria are selected from the group consisting of Klebsiella pneumoniae,   Diplococcus pneumoniae,   Streptococcus pyogenes, and   Hemophilus influenzae.